Biomarker Approach to Screening for the early detection of HPV-related Oropharyngeal Cancer (BASH OPC)

摘要 口咽癌（OPC）的发病率显着增加，其中大多数病例（约80%）由人类引起 乳头瘤病毒（HPV）感染类型16。大多数OPC病例诊断较晚，需要强化治疗。 化学辐射导致严重的发病率、终身残疾和死亡率。早期OPC检测目前 最可行的二级预防方法，有可能大大改善OPC患者的结局。 我们完成了一系列研究，重点是开发一种适用于早期OPC的口腔漱口生物标志物面板 可以在单个口腔漱口液样本中测量的检测，并且可以区分早期OPC（T1-T4）和早期OPC（T2-T4）。 2例N 0 -1 [有单个同侧淋巴结≤3 cm的小肿瘤]）病例和对照。 在一项后续研究中，使用基因组-DNA的生物标志物组特异性和灵敏度得到改善（AUC = 0.935）。 广泛甲基化阵列发现方法。这导致小组扩大到包括口服HPV 16状态， 13个差异甲基化的宿主基因CpG位点。我们现在准备推进这个口腔漱口生物标志物小组 生物标志物开发的第二阶段。我们建议利用现有的早期OPC生物储存库 两个癌症中心的病例和研究基础设施，以有效地推进临床检测开发， 验证。我们的目标是验证一种非侵入性诊断生物标志物面板，以早期检测OPC。中央 假设是，我们可以区分早期OPC病例与无癌个体使用相关的生物标志物， HPV和宿主表观遗传学改变，从而鉴定可以有效和安全地用抗HPV药物治疗的肿瘤。 单一治疗模式下生存率最高。由于我们清楚地意识到这是一个不断发展的领域， 随着文献的发展，对这些和其他生物标志物进行初步评估。主要目的是 评估HPV 16 DNA和宿主基因甲基化联合口腔生物标志物的敏感性和特异性 在100例早期和100例晚期疾病治疗前OPC中区分早期OPC病例和对照组 来自Moffitt癌症中心和UPMC的200例病例和200例对照，这些对照按性别、年龄、种族和烟草进行匹配。 希尔曼癌症中心次要目的包括评估实验室间和实验室内生物标志物的一致性 和可靠性，分别在病例和对照组中与生物标志物组相关的因素， 其它差异甲基化的宿主基因（例如，SYNGR 3）和/或口腔肿瘤组织修饰的HPV DNA检测早期OPC。这是一种创新的OPC早期检测方法，使用非侵入性标本 容易获得（例如，在常规牙科实践中）。该方法建立在现有的临床/流行病学 基础设施、注释良好的生物储存库和来自OPC病例和对照的数据以及强有力的初步数据。 我们的研究团队拥有成功实施拟议研究的经验和专业知识。结果将 为在非侵入性研究中测量的生物标志物面板的大型确定性前瞻性筛选试验的设计提供信息。 在高风险个体中检测样本，以将OPC诊断从晚期转移到早期疾病。

ABSTRACT Oropharyngeal cancer (OPC) incidence is significantly increasing with most cases (~80%) caused by human papillomavirus (HPV) infection type 16. The majority of OPC cases are diagnosed late and require intensive chemo-radiation causing significant morbidity, life-long disabilities, and mortality. Early OPC detection is currently the most viable secondary prevention approach with the potential to vastly improve outcomes for OPC patients. We completed a series of studies focused on developing an oral gargle biomarker panel suitable for early OPC detection that can be measured in a single oral gargle specimen, and can discriminate between early OPC (T1- 2 N0-1 [small tumors with a single ipsilateral node ≤3 cm]) cases and controls in two different case-control sets. In a follow-up study the biomarker panel specificity and sensitivity was improved (AUC = 0.935) with a genome- wide methylation array discovery approach. This resulted in panel expansion to include oral HPV 16 status and 13 differentially methylated host gene CpG sites. We are now poised to advance this oral gargle biomarker panel to the second phase of biomarker development. We propose to leverage existing biorepositories of early OPC cases and research infrastructure at two cancer centers to efficiently advance clinical assay development and validation. Our goal is to validate a non-invasive diagnostic biomarker panel to detect OPC early. The central hypothesis is that we can distinguish early OPC cases from cancer-free individuals using biomarkers related to HPV and host epigenetic alterations, thereby identifying tumors that can be effectively and safely treated with a single modality where survival is highest. As we are keenly aware that this is an evolving field, we will also conduct preliminary evaluation of these and other biomarkers as the literature evolves. The Primary Aim is to estimate the sensitivity and specificity of a combined HPV 16 DNA and host gene methylation oral biomarker panel to distinguish early OPC cases from controls among 100 early and 100 late disease pre-treatment OPC cases, and 200 controls matched by sex, age, race, and tobacco from the Moffitt Cancer Center and the UPMC Hillman Cancer Center. Secondary Aims include assessing inter- and within-laboratory biomarker concordance and reliability, factors associated with the biomarker panel separately among cases and controls, and the performance of other differentially methylated host genes (e.g., SYNGR3) and/or oral tumor-tissue modified HPV DNA to detect OPC early. This is an innovative approach to OPC early detection, using a non-invasive specimen readily obtained (e.g., in routine dental practice). The approach builds on existing clinical/epidemiologic infrastructure, well annotated biorepository and data from OPC cases and controls, and strong preliminary data. Our research team has the experience and expertise to successfully implement the proposed study. Results will inform the design of a large definitive prospective screening trial of a biomarker panel measured in a non-invasive specimen among high-risk individuals to shift OPC diagnosis from late to early disease.